摘要
为了快速有效地确定泡桐属植物分子发育系统PCR反应体系,将影响PCR扩增体系的主要单因素因子Taq DNA聚合酶、Mg^(2+)、dNTPs替换成Taq PCR Master Mix;通过模板DNA,引物浓度和退火温度对体系进行优化,建立最适合泡桐属植物叶绿体DNA的PCR最适反应体系(25μL):5μL模板DNA,1.0μL引物,12.5μL Taq PCR Master Mix,5.5μL ddH_2O,反应体系具有较高的稳定性和重复性;筛选出了适合泡桐属植物分子系统发育学研究的引物:trb V-ndh C,Agt1F-Agt1R,ITS4-ITS5.
In order to quickly and effectively identify the species of the genus paulownia molecular PCR reactionsystem,Taq PCR Master Mix replaces Taq DNA polymerase,d NTPs and Mg^（2＋） which effect the system of PCRamplification to establish and optimization of the system through template DNA,primer concentrations and annealingtemperature to bulid the most suitable plants of the genus paulownia chloroplast DNA PCR optimal reaction systemof 25 μL,as follows：5 μL template DNA,1.0 μL primers,12.5 μL Taq PCR Master Mix,5.5 μL ddH_2O,the reactionsystem has high stability and repeatability,to select the primers which suitable the plants of the genus paulowniamolecular phylogenetic studies：trb V-ndh C,Agt1F-Agt1 R,ITS4-ITS5.
出处
《河南科学》
2016年第2期208-213,共6页
Henan Science
基金
国家科技支撑计划专题(2013BAD01B06)
郑州市科技领军人才计划(10LRC177)
